Materials and Reagents

1. Plant material
   1. 4 to 5 week old healthy N. benthamiana plants
      
   
   
2. Vectors and bacteria strains
   1. pDONR221 or other gateway DONR vector (Life Technologies, Invitrogen^TM, catalog number: 12536-017 )
      
   2. Escherichia coli (E. coli) DH10B or similar cells for molecular cloning
      
   3. SGR reporter destination vector, SPDK 2388
      Note: This vector contains the reporter gene SGR without promoter.
      
   4. Binary vector for TFs over-expression (e.g. LIC6 from Arabidopsis Biological Resource Center)
      
   5. Binary vector for expression of negative control protein [e.g. Actin7 (At5g09810) in LIC6]
      
   6. Agrobacterium tumefaciens (A. tumefaciens) strain GV2260
      
   
   
3. Other materials
   1. BP and LR clonase enzyme Kit (Life Technologies, Invitrogen^TM, catalog numbers: 11789-013 and 11791-043 )
      
   2. LB media
      
   3. Acetosyringone (Sigma-Aldrich, catalog number: D134406 )
      
   4. Infiltration medium (see Recipes)
      

Equipment

1. Incubator (42 °C)
   
2. 1 ml Tuberculin syringes without needle (Tyco, catalog number: 8881501400 )
   
3. Controlled environment plant growth chamber (12 h of light per day, 21 °C)
   

Procedure